Pharmaceutical and/or cosmetic composition containing an active antioxydant principle and cell energy activator

ABSTRACT

A cosmetic or pharmaceutical composition, and particularly dermatological composition, comprising, in a physiologically adapted medium, as active principle, polypeptides or peptides, and methods of administering a composition capable of increasing cell energy and to protect the skin from oxidative damage. Also, a cosmetic treatment procedure aimed at protecting the skin and the skin appendages from external aggressions and to combat skin aging.

The present invention is in the cosmetic and pharmaceutical field, andmore specifically in the field of dermatology. The present inventionconcerns a cosmetic or pharmaceutical composition, and particularlydermatological, comprising, in a physiologically adapted medium, asactive principle, peptides, used alone or in association with at leastone other active principle. The invention also relates to the use of acomposition capable of increasing cell energy and to protect the skinfrom oxidative damage. The invention also relates to a cosmetictreatment procedure aimed to protect the skin and skin appendages fromexternal aggressions and to combat skin aging. The active principle canalso be used to prepare pharmaceutical compositions aimed at preventingor combating pathologies linked to oxidation processes or also, certainaging pathologies.

According to the invention, the term “skin appendages” includes allkeratin annexes present on the surface of the body, in particular thehair, the lashes, the eyebrows, the nails and hair.

The skin is a vital organ which covers the entire surface of the bodyand insures the protective, sensitive, immune, metabolic or alsothermoregulation functions. The skin, like the other organs, is subjectto aging. Now, one of the major mechanisms involved in aging processesis the accumulation of oxidative damage in essential molecules such asmembrane lipids, proteins, ADN and quite particularly mitochondrial ADN(ADNmt).

Oxidative damage is caused by free radicals, chemically instable andhighly reactive molecules, generated by intracellular metabolism orexternal aggressions. External aggressions include: UV rays, toxins,atmospheric pollutants, food oxidants. In the skin, one observespremature aging manifested in those areas exposed to rays, characterizedby macromolecule alteration phenomena (lipid peroxydation, carbonylationof proteins) affecting in particular elastin, collagen or fibronectine.One has also been able to show/demonstrate a progressive decline in themitochondrial functions with age, probably linked to the accumulation ofmutations to ADNmt (K. Singh, Ann. N.Y. Acad. Sci. 1019, 2004).

One of the most important consequences of the accumulation of oxidativedamage is a reduction of the cell's capacity to produce ATP (Porteous etal., Eur J Biochem 1998, 257(1):192-201). Thus, the phenomenon ofcellular aging concerns the oxidative damage which the cellsuffers/undergoes but also with the energy production processes neededfor the cell necessary to survive.

The body possesses defense mechanisms capable of trapping ortransforming free radicals (enzymes, glutathione, vitamins A and E,coenzyme Q10, etc.). However, these antioxidant defense systems areoften insufficient against the many stress and external aggressions towhich the organisms and skin in particular are subjected.

In this context, the antioxidant properties of the coenzyme Q10 appearto be particularly interesting:

Coenzyme Q10 (or ubiquinone) is a coenzyme present in the mitochondrialcomplexes involved in the oxidative phosphorylation lead to theproduction of ATP (Mitchell et al, 1976; Mitchell et al, 1990). Theother fundamental property of the coenzyme Q10 is being an antioxidant,neutralizing free radicals (Beyer et al 1990, Villalba et al 1997).

Coenzyme Q10 is a benzoquinone derivative flanked by a long lateralisoprene chain consisting more often of ten isoprenoid units (hence thename Coenzyme Q10). As this coenzyme is not water soluble, it is onlyfound in lipid membranes such as the internal membrane of themitochondria, where it can freely be released from the phospholipidmembranes.

Coenzyme Q10 can exist in three states of oxidation: a reduced form(CoQH2 or UQH2), an oxidized form (CoQ10), and an intermediary form theradical ubisemiquinone (Q).

Coenzyme Q10 is present in the skin where it stimulates natural cellfunctions and acts to defend against external aggressions.

The biosynthesis of coenzyme Q10 is done by starting with the tyrosinefor the quinone kernel, and starting with farnesyl pyrophosphate for thelateral chain. The enzyme responsible for this last reaction, which isan essential stage in the biosynthesis of the coenzyme Q10, istransprenyl transferase (or polyprenyl transferase).

The search for compositions able to stimulate the synthesis of coenzymeQ10, the energy synthesis of ATP and/or to protect cells damage causedby free radicals is a concern of medical and cosmetic research.Solutions has therefore been proposed to use peptide substances havingantioxidant properties (WO2005097060, JP2006131626), but any cosmetic orpharmaceutical composition comprising the peptides or polypeptides ofthe present invention has yet to be described.

The inventors have shown a therapeutic activity, and more particularlydermatological and cosmetic activity, special peptides, described in thepresent invention. It has particularly been demonstrated that thesepeptides, when applied to the skin, have a strong protective activitywith respect to the oxidative damage suffered by the skin and favoringto a significant degree the synthesis of ATP, and the synthesis or theactivity of the enzyme transprenyl transferase and the coenzyme Q10.This new active principle, capable of increasing cell energy andprotecting the skin from oxidative damage, thus makes it possible toopen up new therapeutic and cosmetic perspectives.

“Active principle capable of increasing cell energy and protecting theskin from oxidative damages” means all substances capable of increasingthe synthesis of intracellular ATP and to provide protective propertiesin the cells or tissues subject to physical-chemical or environmentaloxidant stress.

Thus, the first purpose of the invention is a cosmetic or pharmaceuticalcomposition, and particularly dermatological, comprising, in aphysiologically adapted medium, as active principle, peptides capable ofincreasing cell energy and protecting the skin from oxidative damage,alone or in association with at least one other active principle.

The term “peptide” indicates a chain of two or more amino acids linkedby peptide bonds or by modified peptide bonds.

“Peptide” means the natural or synthetic peptide according to theinvention as described above or at least one of its fragments, which areobtained by proteolysis or synthetically, or also all natural orsynthetic peptides in which the sequence is totally or partiallycomprised by the sequence of the peptide according to the invention.

Preferably, according to this invention, the active principle accordingto the invention or its biologically active derivatives is compositionof at least one peptide whose number of amino acids is between 5 and 13.

The expression “biologically active” means “that which possesses an invivo or in vitro activity characteristic of the activity of the activeprinciple according to the invention.”

According to a particular embodiment of the invention, the peptidepossesses a sequence that corresponds to the general formula (I)

R₁-(AA)_(n)-Ala-Val-Leu-Ala-Gly-(AA)_(p)-R₂

In which

AA represents any amino acid, or one of its derivatives, and n and p arewhole numbers between 0 and 4.

R₁ represents the primary amino function of the amino acid N-terminal,free or substituted by a protector group which can be selected from anacetyl group, a benzyl group, a tosyl group or a benzyloxycarbonylgroup.

R₂ represents the hydroxyl function of the carboxyl acid of C-terminalamino acid, free or replaced by a protector group which can be selectedfrom an alkyl chain from C1 to C₂₀, or a NH₂, NHY or NYY group with Yrepresenting an alkyl chain from C1 to C₄.

According to a particularly preferred embodiment of the invention, thebiologically active peptide is the sequence SEQ ID no 1 or the sequenceSEQ ID no 2

(SEQ ID n^(o) 1) Ala-Val-Leu-Ala-Gly-Asp-NH2 (SEQ ID n^(o) 2)Ala-Val-Leu-Ala-Gly-Asp

The invention also concerns homologous forms of these sequences. Theterm “homologous” designates, according to the invention, all peptidesequences at least 50% identical, or preferably at least 80%, and alsomore preferably at least 90% identical to the referenced peptidesequence, selected between the sequence SEQ ID no 1 and the sequence SEQID no 2. “at least X % identical Peptide sequence” designates a percentof identity between the residues of amino acids of the two sequences tobe compared, obtained after the optimal alignment of the two sequences.The optimal alignment is obtained using algorithms of local homologoussuch as those used by the BLAST P or T BLAST N software available on theNCBI site.

The term “homologous” can also indicate a peptide which differs from thesequence of a peptide from the sequence SEQ ID no 1 or SEQ ID no 2 bythe substitution of chemically equivalent amino acids, that it, by thereplacement of one residue by another having the same characteristics.Thus, the classical substitutions are between Ala, Val, Leu and Ile;between Ser and Thr; between the acid residues Asp and Glu; between Asnand Gln; and between the basic residues Lys and Arg; or between thearomatic residues Phe and Tyr.

In the invention, the term “amino acid” refers to all natural ornon-natural organic acids having the formula:

—NHR—CR—C(O)—O—

where each —R is independently chosen between one hydrogen and one alkylgroup having between 1 and 12 carbon atoms. Preferably, at least one —Rgroup of each amino acid is a hydrogen. The term “alkyl” means a carbonchain that can be linear or branched, substituted (mono- or poly-) ornon-substituted; saturated, monosaturated (a double or triple bond inthe chain) or polyunsaturated (two or more doubles bonds, two or moretriples bonds, one or more double bonds and one or more triple bonds inthe chain).

“Peptide” means the natural or synthetic peptide according to theinvention as described above or at least one of its fragments, which isobtained by proteolysis or synthetically, or also all natural orsynthetic peptides whose sequence is totally or partially constituted bythe sequence of the peptide described above.

In order to improve resistance to degradation, it may be necessary touse a protected form of the peptide according to the invention. The formof protection must evidently be a biologically compatible form and mustbe compatible with use in the field of cosmetics or pharmacy.

Many forms of biologically compatible protection can be imagined. Thus,the invention concerns a composition as defined above, characterized bythe fact that the general formula peptide (I) possesses at least onefunctional group protected by a protector group, this protector groupbeing either an acylation or an acetylation of the amino-terminal end,or an amidation or an esterification of the carboxy terminal end, or thetwo. The amino-terminal end can be protected by an acetyl group, abenzyl group, a tosyl group or a benzyloxycarbonyl group. Preferably,one uses a protection based on the amidation of the hydroxyl function ofcarboxy terminal end by a NYY group with Y representing an alkyl chainfrom C1 to C₄, or an esterification by an alkyl group. It is alsopossible to protect the two ends of the peptide.

The peptide derivatives also concern the amino acids and the peptideslinked by a pseudo-peptide bond. “Pseudo-peptide bond” means all typesof bonds able to replace the “classical” peptide bonds.

In the field of amino acids, the geometry of the molecules is such thatthey can theoretically take the form of different optic isomers. Thereexists, en effect, a molecular conformation of amino acid (AA) as itturns to the right the polarization plan of the light (dextrorotatoryconformation or D-aa), and a molecular conformation of amino acid (aa)as it turns to the left the light polarization plan (levorotatoryconformation or L-aa). Nature only provides for natural amino acids andlevorotatory conformation. Consequently, a natural origin peptide canonly comprise amino acids of the type L-aa. However, chemical synthesisin the laboratory makes it possible to prepare amino acids having thetwo possible conformations. Starting with this base material, it is thusalso possible during peptide synthesis to incorporate equally well aminoacids in the form of dextrorotatory optical isomers or levorotatory.Thus, amino acids making up the peptide according to the invention canbe under the configuration L- and D-; preferentially, the amino acidsare under form L. The peptide according to the invention can then beunder the form L-, D- or DL-.

The general formula peptide (I) according to the invention can beobtained either by classical chemical synthesis (en solid phase or inhomogenous liquid phase), or by enzyme synthesis (Kullman et al., J.Biol. Chem. 1980, 225, 8234), starting with constituent amino acids ortheir derivatives.

The peptide according to the invention can also be obtained fromfermentation of a strain of bacteria modified or unmodified, by geneticengineering, or also by the extraction of animal or vegetable protein,preferably of vegetable origin, following by controlled hydrolysis whichfrees peptide fragments corresponding totally or partially to thegeneral formula peptides (I).

Many proteins found in plants are likely to contain these sequenceswithin their structure. Controlled hydrolysis makes it possible torelease these peptide fragments. It is possible, but not necessary torealize the invention, to extract both the proteins concerned initiallyand then to hydrolyze them, and to do the hydrolysis initially on a rawextract and then to purify the peptide fragments. It is also possible touse certain hydrolyzed extracts without purifying the peptide fragmentscorresponding to the general formula peptides (I) according to theinvention, but insuring the presence of said fragments by theappropriate analytical means.

Other simpler or more complex procedures can be envisioned by the personskilled in the art familiar with the synthesis, extraction andpurification of proteins and peptides. Thus, the peptide according tothe invention can be of natural or synthetic origin. Preferablyaccording to the invention, the peptide is obtained by chemicalsynthesis. According to the invention, the active principle can be amixture of peptide derivatives and/or made up of amino acid derivatives.

According to an advantageous embodiment of the invention, the activeprinciple according to the invention is made soluble beforehand in oneor more solvents classically used by the person skilled in the art, suchas water, glycerol, ethanol, propylene glycol, butylene glycol,dipropylene glycol, ethoxylated or propoxylated diglycols, cyclicpolyols, Vaseline, a vegetable oil or any mixture of these solvents.

According to still another advantageous embodiment of the invention, theactive principle according to the invention is beforehand made solublein a cosmetic or pharmaceutical vector such as liposomes or adsorbed onorganic powder polymers, mineral supports such as talcs and bentonites,and more generally made soluble in, or fixed on, all cosmetically orpharmaceutically acceptable vectors.

It is well understood that the active principle according to theinvention can be used alone or in association with at least one otheractive principle, in a cosmetic composition or for the preparation of apharmaceutical and/or dermatological composition.

The compositions according to the invention can be applied by allappropriate paths, particularly orally, parenteral or topical external,and their formulation will be adapted by the person skilled in the art,in particular for cosmetic or dermatological compositions.Advantageously, the compositions according to the invention take a formadapted to topical application. The compositions must then contain acosmetically and/or dermatologically acceptable medium, that it,compatible with the skin and the skin appendages, and cover all cosmeticor dermatological forms. These compositions can particularly be in theform of creams, oil-in-water emulsions, or water-in-oil or multipleemulsions, solutions, suspensions, gels, milks, lotions, sticks or alsopowders, adapted to application to the skin, the lips and/or the skinappendages.

These compositions comprise the excipients necessary for theirformulation, such as solvents, thickeners, thinners, surfactants,anti-oxidants, dyes, preservatives, perfumes.

To be sure, the person skilled in the art will make sure to selectpossible complementary compositions, active or non-active, and/or theirquantity, in such a way that the advantageous properties of the mixtureare not, or not significantly, altered by the planned addition.

The composition utilizable according to the invention can in particularconsist of a hair care composition, and particularly a shampoo, anafter-shampoo, a waving lotion, a medicated lotion, a cream or a hairgel, a restructuring lotion for hair, a mask, etc. The cosmeticcomposition according to the invention can be used particularly intreatments implementing an application which is followed or not followedby rinsing, or also in the form of shampooing.

It can also take the form of a tincture or mascara to be applied to thebrush or to the comb, in particular on the lashes, eyebrows or hair.

Advantageously, the utilizable compositions also contain at least oneother active agent promoting the action of the peptides according to theinvention. Thus, the composition can associate, to the active principleaccording to the invention, active agents having an antioxidant action,or also stimulating the synthesis of dermal macromolecules, or alsostimulating energy metabolism. For example, as active agents having anantioxidant action, one can cite vitamin C, vitamin E, or polyphenolplant extracts.

One can also cite, as active agents stimulating the syntheses of dermalmacromolecules (laminine, fibronectine, collagen), for example thecollagen peptide marketed under the name “Collaxyl®” by the Vinciencecompany.

Finally, as active agents stimulating energy metabolism, one can citethe active principle marketed under the name “GP4G®” by the Vinciencecompany.According to another aspect, the composition according to the inventioncan be a sun composition, that is, a composition helping to protectagainst sun rays. Thus, it can be advantageously added, to thecomposition according to the invention, active principles used for sunprotection such as, for example, sun blocks.It is quite evident that the invention is directed to mammals ingeneral, and more particularly to human beings.

The effective quantity of the active principle corresponds to thepeptide quantity according to the invention necessary to obtain thedesired result, that is: increase the synthesis of ATP, protect the skinfrom oxidative damage and more generally, protect the skin from externalaggressions and prevent or treat aging skin.

According to an advantageous embodiment of the invention, the generalformula active principle (I) is present in the composition according tothe invention in a concentration between approximately 0.0005 and 500ppm (parts per million), and preferably at a concentration betweenapproximately 0.01 and 5 ppm compared to the total weight of the finalcomposition.

These compositions can particularly be present in the form of an aqueoussolution, hydroalcoholic or oily; of an oil-in-water emulsion,water-in-oil or multiple emulsions; they can also take the form ofcreams, suspensions, or also powders, adapted to an application on theskin, the mucosa, the lips and/or the skin appendages. Thesecompositions can be more or less liquid and have the appearance of acream, a lotion, a milk, a serum, a pomade, a gel, a paste or a mousse.They can also take solid form, such as a stick or be applied to the skinunder aerosol form. They can be used as a care product and/or as a skinmakeup product.

These compositions comprise, in addition, all additives commonly used inthe considered field of application as well as the additives necessaryfor their formulation, such as solvents, thickeners, thinners,antioxidants, dyes, sunblocks, self-tanning [principles], pigments,charges, preservatives, perfumes, air fresheners, cosmetic orpharmaceutical active [principles], essential oils, vitamins, essentialsurfactant fatty acids, polymers leaving a protective film, etc.

In any case, the person skilled in the art will make sure to that theseadditives and their proportions are selected in such a way as not toharm the advantageous properties sought in the composition according tothe invention. These additives can, for example, correspond to from 0.01to 20% of the total weight of the composition. When the compositionaccording to the invention is an emulsion, the fat phase can representfrom 5 to 80% by weight and preferably 5 to 50% by weight compared tothe total weight of the composition. The emulsifiers and co-emulsifiersused in the composition will be selected from those classically used inthe field in question. For example, they can be used in a proportion ofbetween 0.3 to 30% by weight, compared to the total weight of thecomposition.

Due to these special activities, the active principle according to theinvention can be used advantageously in a cosmetic composition or forthe preparation of a pharmaceutical composition.

In particular, the active principle according to the invention can beused advantageously in a cosmetic composition to increase the synthesisof intracellular ATP of skin cells.

The active principle according to the invention can also be usedadvantageously in a cosmetic composition to increase the activity or thesynthesis of the enzyme transprenyl transferase and/or the coenzyme Q10in skin cells.

An essential aspect of the invention is the use of the invention activeprinciple, in a cosmetic composition to protect the skin and the skinappendages from oxidative damage. Said active principle isadvantageously used as antioxidant active principle, and/or asanti-radical active principle, and/or as active anti-glycationprinciple. Anti-radical active principle means all compositions capableof trapping free radicals before the final stages of degradation of thebiologics making up the skin, in that case we refer to antioxidantcompositions. Active anti-glycation principle means any/all compositioncapable of limiting cell damage caused by glycation or glycoxidationreactions. Thus, the active principle according to the invention willmake it possible to combat esthetic damage to the skin and/or haircaused by free radicals.

Also, the active agent can be used advantageously in a cosmeticcomposition to protect the skin and the skin appendages against alltypes of external aggressions. The expression “external aggression”means aggressions that can be produced by the environment. For example,one can cite aggressions such as pollution, UV rays, or also productscreating irritation such as surfactants, preservatives or perfumes.Pollution also means/includes “external” pollution, due for example todiesel particles, ozone or heavy metals, that “internal” pollution thatcan be due particularly to emissions of paint, glue, or wallpaper (suchas astoluene, styrene, xylene or benzaldehyde), or also cigarettesmoking.

Another aspect of the invention is the use of said active principle in acosmetic composition or for the preparation of a pharmaceuticalcomposition, as active photo-protector principle and, more particularly,as active photo-protector principle known as “secondary.” In fact, adistinction is made between primary active photo-protectors principlesand secondary active photo-protectors principles. Primary activephoto-protectors principles are substances which exert a physical power:they are able to absorb UV rays and to return them in the form of heatin order to protect the skin. Secondary active photo-protectorsprinciples are substances which instead have a biological effect; theseare, for example, active antioxidant-type principles which interrupt thechains of photochemical reactions which are triggered when UV rayspenetrate the skin.

In another essential aspect of the invention, the active principle canbe used advantageously in a cosmetic composition to fight in apreventive and/or curative fashion against the signs of skin aging, andmore specifically, to combat and/or prevent light induced aging(photo-aging). Skin aging signs means all modifications of the exteriorappearance of the skin and the skin appendages due to aging such as, forexample, lines and wrinkles, wilted skin, flabby skin, drawn skin, lackof elasticity and/or skin tone, dull skin or skin pigment spots, hairdiscoloration or spots on nails, but also all internal changes of theskin which are not systematically translated by a changed exteriorappearance such as, for example, all oxidative damage of the skinresulting from exposure to ultraviolet rays (UV). The active principleaccording to the invention, or the composition containing it, will makeit possible to fight, in particular, against loss of skin elasticity andfirmness.

The purpose of the invention is also to be used in a cosmeticcomposition of an effective quantity of active principle as describedabove, to prevent the damage caused to the skin by exposure to the sunor exposure to ionizing rays during radiotherapy.

The purpose of the invention is also the use in a cosmetic composition,of an effective quantity of active principle as described above, tostimulate and/or protect mitochondria, in particular on the areas of thebody exposed to UV rays.

The invention also consists of a pharmaceutical compositioncharacterized in that the active principle according to the invention isformulated to alleviate a pathology linked to oxidation processes, oralso certain aging pathologies.

The invention also consists of a cosmetic treatment procedure aimed atprotecting the skin and the skin appendages from external aggressionsand to fight skin aging characterized by the application to the skin orthe skin appendages to be treated a composition containing an effectivequantity of active principle according to the invention.

Special embodiments of this cosmetic treatment procedure are also shownin the description above. Other advantages and characteristics of theinvention will be more apparent by reading examples provided fornon-limiting illustrative purposes.

EXAMPLE 1 Assessment of the Activator Effect of the Peptide SEQ ID No 2on the Synthesis of Intracellular ATP

The goal of this study is to determine the influence of the peptide SEQID no 2 on the synthesis of ATP.

Protocol

This study was conducted using an “ATP Bioluminescence Assay Kit HS II”Kit (Roche Applied Science):

The dermal fibroblasts are treated with a 1% solution, a solution a 50ppm, containing the peptide SEQ ID no 2, representative of the family ofpeptides according to the invention, for a period lasting from 1 to 3hours. At the end of the incubation period, the wells are emptied oftheir medium and rinsed with 2 ml of cold PBS before adding 250 μl of alysis buffer supplied by the kit. The cells of each well are thenscrapped, then collected in 14 ml tubes. Each well is rinsed with 2×500μl of cold PBS and everything is against collected the respective tubes.Beginning with these samples, a 1/12000^(th) dilution is done in coldPBS before each reading. The dosage of ATP is done on these samples: 50μL of this dilution is placed in a luma basin and 50 μl of luminol areadded. After 10 seconds, the luminescence read is triggered. The valuesare standardized with respect to the quantity of proteins for eachsample. The measures are done using an apparatus: the Biocounter M2010ALUMAC®/3M.

Results

The measurements of ATP show that there is a 17% increase in thequantity of intracellular ATP after 1 hour and 67% after 3 hours ofculture in cells treated by the peptide SEQ ID No 2, compared to theuntreated cells.

Conclusion

The peptide SEQ ID no 2 greatly activates the synthesis of intracellularATP in skin cells.

EXAMPLE 2 Assessment of the Protector Effect of the Peptide SEQ ID No 1with Respect to Oxidative Damage

The goal of this study is to determine the protector effect of thepeptide SEQ ID no 1 with respect to dermal fibroblasts subjected tooxidative stress caused by UV rays or hydrogen peroxide (H₂O₂). Toevaluate the oxidative damage suffered by the cells, measurements of thecarbonylation of the proteins was done.

The carbonylation of proteins is the result of the oxidative splittingof proteins or oxidation of the arginine residues, lysine, proline orthreonine. The dosage of the carbonylation of proteins is done using anEIA (Enzyme Immuno Assay) technique.

Protocol

Fibroblasts in culture were placed in the presence of the peptide SEQ IDno 1 in a 1% solution, at 50 ppm, 72 hours before, during, and also 24hours after the oxidative stress (UVB irradiation of 50 mJ/cm² ortreatment par 2 mM of H₂O₂). untreated controls not subjected tooxidative stress are done.

The degree of carbonylation consists in the use of DNP (dinitrophenyl)which has the property to se fixer specifically on these carbonyl groupsof the proteins. The fixed DNP is then assayed using an ELISA method,thanks to an anti-DNP antibody linked to peroxydase. A range of BSA(bovine albumin serum) oxidized (with a known concentration of carbonylgroups) is used for calibration.

Results

The results achieved show a 30% reduction in the carbonylation ofproteins when the cells are treated with the peptide SEQ ID no 1according to the invention, compared to the untreated cells.

More particularly, one observes a 20% decrease in carbonylation when thecells treated with the peptide SEQ ID no 1 are subjected to irradiationby UVB or to an oxidative stress by H₂O₂, compared to the irradiated orstressed cells not treated with the active principle.

Conclusion

The peptide SEQ ID no 1 effectively protects the skin cells againstoxidative damage caused by UVB rays or hydrogen peroxide.

EXAMPLE 3 Assessment of the Protector Effect of the Peptide SEQ ID No 1with Respect to Stress Induced by Glycation

The goal of this study is to determine the protective effective of thepeptide SEQ ID no 1, with respect to ex vivo epidermal culture subjectedto stress by a glycation agent.

Protocol

Biopsies of human skin are maintained in ex vivo culture, treated with a1% solution, a mother solution of 50 ppm of peptide SEQ ID no 1, 24hours before, and also 24 hours after the introduction of a glycationagent (methyl glyoxal at 5 or 10 mM). Histology hematoxyline-eosine(H&E) staining makes it possible to evaluate the quality of skinstructures.

Results

The evaluation of the skin structures shows a net resistance to cellularstress induced by glycation of biopsies of skin treated with the peptideSEQ ID no 1.

Conclusion

The peptide SEQ ID no 1 protects the skin from stress induced byglycation.

EXAMPLE 4 Preparation of Compositions

1—Sun Protection Cream:

Trade names INCI Names % W/W PHASE A Demineralized water Aqua (Water)qsp Pemulen TR1 Acrylates/C10-30 Alkyl Acrylate 0.40 CrosspolymerGlycerin Glycerin 3.00 Nipastat Sodium Sodium Methylparaben (and) Sodium0.15 Ethylparaben (and) Sodium Butyl paraben (and) Sodium Propylparaben(and) Sodium Isobutylparaben PHASE B Parsol MCX EthylhexylMethoxycinnamate 7.50 Eusolex 4360 Benzophenone-3 3.00 Parsol 1789 ButylMethoxydibenzoylmethane 2.00 Myritol 318 Caprylic/Capric Triglyceride4.00 Emulgade SEV Hydrogenated Palm Glycerides (and) 5.00 Ceteareth-20(and) Ceteareth-12 (and) Cetearyl Alcohol Propylparaben Propylparaben0.15 Nacol 16-98 Cetyl Alcohol 1.00 PHASE C TEA Triethanolamine 0.20PHASE D Peptide SEQ ID n^(o) 1 3 ppm Perfume Perfume (Fragrance) qsp DyeqspThe components of phase A and phase B are heated separately to between70° C. and 75° C. Phase B is emulsified in phase A under agitation.Phase C is added, to 45° C., increasing the agitation. Phase D is thenadded when the temperature is below 40° C. The cooling is continued to25° C. under lively agitation.

2—After-Sun Milk:

Trade names INCI Names % w/w PHASE A Montanov L C14-22 Alcohols (and)C12-20 3.00 Alkyl Glucoside Waglinol 2559 Cetearyl Isononanoate 4.00Tegosoft TN C12-15 Alkyl Benzoate 3.00 Apricot kernel oil PrunusArmeniaca (Apricot) 2.00 Kernel Oil Avocado oil Persea Gratissima(Avocado) Oil 1.00 Abil 350 Dimethicone 1.00 PHASE B Demineralized WaterAqua (Water) qsp PHASE C Simulgel EG Sodium 0.4 Acrylate/Acryloyldimethyl Taurate Copolymer (and) Isohexadecane (and)Polysorbate 80 Copolymer (and) Polysorbate 80 PHASE D PhenonipPhenoxyethanol (and) 0.30 Methylparaben (and) Ethylparaben (and)Butylparaben (and) Propylparaben (and) Isobutylparaben Ethylparaben andPropylparaben and Buthylparaben Germall 115 Imidazolidinyl Urea 0.20PHASE E Peptide SEQ ID n^(o) 2 0.1 ppmPrepare phase A under agitation. Progressively incorporate the xanthangum, under deflocculating agitation. Phases C and D will be incorporatedonce the gel is ended. Phase E, prepared beforehand until the DHA isfully dissolved, will then be added. Adjust the pH if necessary to4-4.5. Dye and perfume.

3—Anti-Aging Cream:

Trade names INCI Names % w/w Phase A Montanov 68 Cetearyl Alcohol (and)Cetearyl Glucoside 6.00 Squalane Squalane 3.00 Cetiol SB 45Butyrospermum Parkii (Shea Butter) 2.00 Waglinol 250 CetearylEthylhexanoate 3.00 Amerchol L-101 Mineral Oil (and) Lanolin Alcohol2.00 Abil 350 Dimethicone 1.50 BHT BHT 0.01 Phase B Avocado oil PerseaGratissima (Avocado) Oil 1.25 Phenonip Phenoxyethanol (and)Methylparaben 0.75 (and) Ethylparaben (and) Butylparaben (and)Propylparaben (and) Isobutylparaben Phase C Demineralized water Aqua(Water) qsp Butylene Glycol Butylene Glycol 2.00 Glucam E10 MethylGluceth-10 1.00 Allantoin Allantoin 0.15 Carbopol Ultrez 10 Carbomer0.20 Phase D TEA Triethanolamine 0.18 Phase E Peptide SEQ ID n^(o)1 0.5ppm GP4G Water (and) Artemia Extract 1.50 Collaxyl Water (and) ButyleneGlycol (and) 3.00 Hexapeptide-9 Phase F Perfume Perfume (Fragrance) qspDye qspPrepare and blend phase A at 65-70° C. Heat phase C at 65-70° C. Phase Bis added at phase A just before emulsifying A in B. A approximately 45°C., the carbomer is neutralized by addition of phase D. Phase E is thenadded under light agitation and cooling is continued to 25° C. Phase Fis then added if desired.

4—Day Protector Cream:

Brand names INCI Name % w/w Phase A Emulium Delta Cetyl alcohol (and)Glyceryl Stearate 4.00 (and) PEG-75 Stearate (and) Ceteth-20 (and)Steareth-20 Lanette O Cetearyl Alcohol 1.50 D C 200 Dimethicone 1.00Fluid/100cs DUB 810C Coco Caprylate/Caprate 1.00 DPPG Propylene GlycolDipelargonate 3.00 DUB DPHCC Dipentaerythrityl Hexacaprylate/ 1.50Hexacaprate Cegesoft PS6 Vegetable Oil 1.00 Vitamin E Tocopherol 0.30Phenonip Phenoxyethanol (and) Methylparaben 0.70 (and) Ethylparaben(and) Butylparaben (and) Propylparaben (and) Isobutylparaben Phase BDemineralized water Aqua qsp 100 Glycerin Glycerin 2.00 Carbopol EDT2020 Acrylates/C10-30Alkyl Acrylate 0.15 Crosspolymer Keltrol BT XanthanGum 0.30 Phase C Sodium Hydroxide Sodium Hydroxide 0.30 (10% solution)Phase D Demineralized water Aqua 5.00 Stay-C 50 Sodium AscorbylPhosphate 0.50 Phase E Butylene Glycol Butylene Glycol 2.00 Dekaben CPChlorphenesin 0.20 Phase F GP4G Water (and) Artemia Extract 1.00 PeptideSEQ ID n^(o)2 5 ppmPrepare phase A and heat to 75° C. under agitation. Prepare phase Bdispersing the carbopol, then the xanthan gum under agitation. Let rest.Heat to 75° C.At temperature, emulsify A in B under rotor-stator agitation. Neutralizewith phase C under rapid agitation. After cooling to 40° C., add phaseD, then phase E. Cooling is continued under light agitation and phase Fadded.

1-18. (canceled)
 19. A composition comprising: an active principle in aphysiologically acceptable medium, said active principle comprising apeptide, or a biologically active derivative thereof, wherein, saidpeptide comprises from 5 to 13 residues of amino acids, and said peptidehas a sequence according to formula (I):R₁-(AA)_(n)-Ala-Val-Leu-Ala-Gly-(AA)_(p)-R₂

in which AA represents any amino acid, or derivative thereof, and n andp are whole number between 0 and 4, R₁ represents the primary aminofunction of the amino acid N-terminal, free or substituted by aprotector group selected from the group consisting of an acetyl group, abenzyl group, a tosyl group and a benzyloxycarbonyl group, R₂ representsthe hydroxyl function of carboxyl acid of the amino acid C-terminal,free or substituted by a protector group selected from (i) an alkylchain from C1 to C₂₀, or (ii) an NH₂ group, an NHY group or an NYY groupwith Y representing an alkyl chain from C1 to C₄.
 20. The compositionaccording to claim 19, wherein said peptide has the sequence SEQ ID No.1 Ala-Val-Leu-Ala-Gly-Asp-NH2.
 21. The composition according to claim19, wherein said peptide has the sequence SEQ ID No. 2Ala-Val-Leu-Ala-Gly-Asp.
 22. The composition according to claim 19,wherein said peptide possesses at least one functional group protectedby a protector group, said protector group being either at least one of(i) an acylation or an acetylation of the amino-terminal end and (ii) anamidation or an esterification of the carboxyterminal end.
 23. Thecomposition according to claim 19, wherein said active principle ispresent in the composition in a concentration between 0.0005 and 500 ppmapproximately compared to the total weight of the composition.
 24. Thecomposition according to claim 23, wherein said active principle ispresent in the composition in a concentration between 0.01 and 5 ppmcompared to the total weight of the composition.
 25. The compositionaccording to claim 19, wherein said active principle made soluble in oneor more solvents cosmetically or pharmaceutically acceptable selectedfrom the group consisting of water, glycerol, ethanol, propylene glycol,butylene glycol, dipropylene glycol, ethoxyl or propoxyl diglycols,cyclic polyols, Vaseline, a vegetable oil and mixtures thereof.
 26. Thecomposition according to claim 19, wherein the composition is in atopical application form.
 27. The composition according to claim 19,further comprising at least one other active agent promoting the actionof said active principle.
 28. The composition according claim 27,wherein said other active agent is selected from the group consisting ofactive agents having an antioxidant action, active agents stimulatingthe synthesis of macromolecules, and active agents stimulating energymetabolism.
 29. A method of preparing a cosmetic composition or apharmaceutical composition according to claim 19, comprising adding aneffective amount of active principle to a physiologically acceptablemedium.
 30. A method of increasing the synthesis of intracellular ATP inthe cells of the skin, comprising administering to a subject in needthereof a cosmetic composition according to claim 19 having an effectiveamount of active principle.
 31. A method of increasing the activity orthe synthesis of the enzyme transprenyl transferase and/or of thecoenzyme Q10 in the cells of the skin, comprising administering to asubject in need thereof a cosmetic composition according to claim 19having an effective amount of active principle.
 32. A method ofprotecting the skin and the skin appendages against all types ofexternal aggressions, comprising administering to a subject in needthereof a cosmetic composition according to claim 19 having an effectiveamount of active principle.
 33. A method of preventing or treatingdamage caused to the skin and the skin appendages by UV rays, comprisingadministering to a subject in need thereof a cosmetic compositionaccording to claim 19 having an effective amount of active principle.34. A method of protecting the skin and the skin appendages fromoxidative damage, comprising administering to a subject in need thereofa cosmetic composition according to claim 19 having an effective amountof active principle.
 35. A method of preventing or treating the skinsigns of aging and/or photoaging, comprising administering to a subjectin need thereof a cosmetic composition according to claim 19 having aneffective amount of active principle.
 36. A method for preparing apharmaceutical composition according to claim 19 for preventing orcombating pathologies linked to the oxidation processes, comprisingadding an effective amount of active principle to a physiologicallyacceptable medium.
 37. A method of treating the skin and the skinappendages from external aggressions and to combat skin aging,comprising topically applying to the skin or the skin appendages of asubject in need thereof a cosmetic composition according to claim 19.